Oligomeric compounds comprising bicyclic nucleosides and uses thereof

a technology of bicyclic nucleosides and compounds, applied in the field ofoligomeric compounds comprising bicyclic nucleosides, can solve the problems of cleavage of mrna and interfere with polyadenylation

Active Publication Date: 2015-09-24
IONIS PHARMA INC
View PDF1 Cites 65 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In certain instances, such binding of an antisense compound to its target mRNA results in cleavage of the mRNA.
Such antisense compounds alter splicing, interfere with polyadenlyation or prevent formation of the 5′-cap of a pre-mRNA.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Oligomeric compounds comprising bicyclic nucleosides and uses thereof
  • Oligomeric compounds comprising bicyclic nucleosides and uses thereof
  • Oligomeric compounds comprising bicyclic nucleosides and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Antisense Inhibition of Human Target X in HuVEC Cells

[0349]Antisense oligonucleotides were designed to target a messenger RNA molecule (Target X) and were tested for their effects on target X mRNA in vitro. Cultured HuVEC cells at a density of 20,000 cells per well were transfected using electroporation with 250 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Target X mRNA levels were measured by quantitative real-time PCR. Target X mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target X, relative to untreated control cells. A total of 40 oligonucleotides were tested. Only those oligonucleotides which were selected for further study are shown in Table 4.

[0350]The ‘Chemistry’ column describes the sugar modifications of each oligonucleotide. ‘k’ indicates an (S)-cEt sugar modification; the number indicates the number of deoxynucleo...

example 2

Dose-Dependent Antisense Inhibition of Human Target X in HuVEC Cells

[0351]Gapmers from the studies described above exhibiting significant in vitro inhibition of Target X mRNA were selected and tested at various doses in HuVEC cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 31.3 μM, 62.5 μM, 125.0 μM, 250.0 μM, 500.0 μM, and 1000.0 μM concentrations of antisense oligonucleotide, as specified in Table 5. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Target X mRNA levels were measured by quantitative real-time PCR. Target X mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target X, relative to untreated control cells. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below.

[0352]T...

example 3

Dose-Dependent Antisense Inhibition of Human Target X in HuVEC Cells

[0353]Additional antisense oligonucleotides were designed as deoxy, MOE and (S)-cEt oligonucleotides targeting Target X gene sequences and were tested at various doses in HuVEC cells. The ‘Chemistry’ column describes the sugar modifications of each oligonucleotide. ‘k’ indicates an (S)-cEt sugar modification; the number indicates the number of deoxynucleosides; otherwise indicates deoxyribose; and ‘e’ indicates a MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate linkages. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in Table 6 is targeted to the human Target X genomic sequence. Oligonucleotides having the same start site and stop site...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
pharmaceutical compositionaaaaaaaaaa
chemical analysisaaaaaaaaaa
chemicalaaaaaaaaaa
Login to view more

Abstract

The present invention provides oligomeric compounds. Certain such oligomeric compounds are useful for hybridizing to a complementary nucleic acid, including but not limited, to nucleic acids in a cell. In certain embodiments, hybridization results in modulation of the amount activity or expression of the target nucleic acid in a cell. In certain embodiments, the present invention provides compounds comprising oligonucleotides. In certain embodiments, such oligonucleotides comprise a region having a gapmer sugar motif. In certain embodiments, oligonucleotides comprise one or more type of modified sugar moieties and / or naturally occurring sugar moieties arranged along an oligonucleotide or region thereof in a defined pattern or sugar modification motif.

Description

BACKGROUND OF THE INVENTION[0001]Antisense compounds have been used to modulate target nucleic acids. Antisense compounds comprising a variety of chemical modifications and motifs have been reported. In certain instances, such compounds are useful as research tools, diagnostic reagents, and as therapeutic agents. In certain instances antisense compounds have been shown to modulate protein expression by binding to a target messenger RNA (mRNA) encoding the protein. In certain instances, such binding of an antisense compound to its target mRNA results in cleavage of the mRNA. Antisense compounds that modulate processing of a pre-mRNA have also been reported. Such antisense compounds alter splicing, interfere with polyadenlyation or prevent formation of the 5′-cap of a pre-mRNA.[0002]Certain antisense compounds have been described previously. See for example U.S. Pat. No. 7,399,845 and published International Patent Application No. WO 2008 / 049085, which are hereby incorporated by refer...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/113
CPCC12N15/113C12N2310/11C12N2310/32C12N2310/323C12N2310/3231C12N2320/30C12N2310/321C12N2310/315C12N2310/34C12N2310/346C12N2310/322C12N15/111C12N2310/341C12N2310/3521C12N2310/3525C12N2320/51
Inventor SETH, PUNIT P.FREIER, SUSAN M.MONIA, BRETT P.SWAYZE, ERIC E.
Owner IONIS PHARMA INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap